Fig. (3) Improved reporter dynamics in the GAL4 hybrid format. HEK293 cells were transiently transfected with vectors in 96-well plates. E2 was used to induce ER; dex was used to induce GR. Cells were harvested 24 hours post induction and assayed for luciferase activity with Dual-Glo^TM Assay System. Luminescence was measured on the GloMax luminometer. Induction = treated Fluc RLU/average untreated Fluc RLU. A) Vectors containing either 5X Gal4 UAS- luc2 or luc2P were transfected. Each reporter was co-transfected with either ERα -LBD or GR-LBD. B) Vectors containing either 5X GAL4 UAS-luc2P or 9X GAL4 UAS-luc2P were transfected. Each reporter was cotransfected with ERα -LBD fused to the GAL4 DBD. C & D) Vectors containing either 5X UAS-luc2 or the 9XUAS-luc2P vectors and either GR or ERα -LBD vectors. 1:3 serial dilutions of each compound were added to the wells.